Imaging experiments (Figs. 3 C , 4 D , 5 A ; S1, A and F–I ; S2 D , and S3, D and F ) were performed with an inverted widefield microscope (Ti Eclipse; Nikon). A 100× Plan-Apochromat (NA 1.4) objective lens (Nikon) was used. Images were captured with a z-step size of 300 nm using a scientific complementary metal-oxide semiconductor camera (Zyla; Andor Technology).
Super-resolution localization experiments of SF–pcMT interactions (Figs. S1 C and S3 L ) and Group 2 SF proteins (Fig. 4 C ) were performed via SIM with the Nikon 3D SIM system (Ti 2 Eclipse). A 100× total internal reflection fluorescence objective (NA 1.45) was used. Images were captured with a complementary metal-oxide semiconductor camera (Orca-Flash 4.0; Hamamatsu) with a z-step size of 300 nm. Raw SIM images were reconstructed by the image stack reconstruction algorithm (Nikon Elements).
Confocal microscopy was performed using an inverted microscope (Ti Eclipse) with a 100× Plan-Apochromat (NA 1.43) objective lens (Nikon) and a swept field confocal scan head with the 35-µm slit mode (Prairie Technologies). Images were captured with a charge-coupled device camera (iXon X3; Andor Technology). Confocal images were also acquired with the A1 confocal laser microscope (Nikon). All images were acquired with Nikon Elements with a z-step size of 300 nm at room temperature (Figs. 1 , 2 , S1 E , and S2 A ).
Super-resolution localization experiments of SF–pcMT interactions (
Confocal microscopy was performed using an inverted microscope (Ti Eclipse) with a 100× Plan-Apochromat (NA 1.43) objective lens (Nikon) and a swept field confocal scan head with the 35-µm slit mode (Prairie Technologies). Images were captured with a charge-coupled device camera (iXon X3; Andor Technology). Confocal images were also acquired with the A1 confocal laser microscope (Nikon). All images were acquired with Nikon Elements with a z-step size of 300 nm at room temperature (